Endocytic Regulation of Intestinal Development

肠道发育的内吞调节

基本信息

批准号：
9262626
负责人：
Jean M Wilson
金额：
$ 41.47万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-02-01 至 2022-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9262626
关键词：
AMOT gene Adult Animals Apical Architecture Binding Biological Models Brush Border CRISPR/Cas technology Cell Differentiation process Cell Line Cell Proliferation Cell membrane Complex Cytoplasmic Tail Data Development Disease Drosophila genus Endocytosis Endosomes Enterocytes Epithelial Epithelial Cells Epithelium Extracellular Domain Genes Goals Growth and Development function Hand Human In Vitro Integral Membrane Protein Intestinal Diseases Intestinal Mucosa Intestines Knock-out Knockout Mice Knowledge Laboratories Maintenance Mammals Mediating Membrane Membrane Glycoproteins Membrane Proteins Microscopy Modeling Monomeric GTP-Binding Proteins Morphogenesis Neonatal Pathway interactions Pattern Play Positioning Attribute Proteins Recruitment Activity Regulation Resolution Role Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Protein Sorting - Cell Movement Testing Tight Junctions Transcription Coactivator Transcriptional Regulation Villus WNT Signaling Pathway Work Zebrafish apical membrane endotubin experimental study human disease insight intestinal epithelium intestinal homeostasis neonate notch protein occludin scaffold therapeutic target tool trafficking

项目摘要

Endocytic regulation of intestinal development The intestinal mucosa is a highly differentiated epithelial barrier that is fundamental for normal function of the intestine. Vesicular trafficking is now recognized as an essential regulator of epithelial polarity, integrity and cell differentiation, and studies in model systems have shown that endocytosis and membrane trafficking are essential for normal ontogenesis of the intestine. During development, the epithelial cells of the intestine go through an intensely endocytic stage that precedes the acquisition of mature intestinal architecture. We identified endotubin (EDTB) as a highly conserved integral membrane protein in the endocytic complex of the developing intestine. In vitro, EDTB regulates tight junction assembly and contact-mediated inhibition of proliferation. In addition, it binds the small GTPase Rab14, which regulates trafficking between endosomes and the apical plasma membrane. However the role of EDTB during development is completely unknown. We have generated an intestinal epithelial cell-specific EDTB knockout mouse and, in Preliminary Data, show that EDTB knockout early in development results in aberrant ontogenesis of the neonatal intestine, including loss of the apical endocytic complex, intracellular accumulation of apical plasma membrane proteins, and an aberrant brush border. We will use this EDTB conditional knockout mouse, together with enteroid cultures and cell lines, to identify the mechanistic basis for these changes in epithelial differentiation. We will define the mechanisms for interaction with tight junction proteins and define the domains of EDTB that mediate its effects. Also, we will analyze the role of Rab14 using intestinal enteroid cultures and human intestinal epithelial cell lines. Finally, we will examine the signaling pathways known to impact intestinal differentiation and define their interaction with EDTB and apical endosomes. Collectively, these studies will provide insight into the cellular mechanisms of intestinal development, which is essential for our understanding of intestinal function. When completed, the experiments outlined in this proposal will define the role of vesicular trafficking in normal growth, development, and homeostasis of the intestinal epithelium. This fundamental knowledge will have implications for our understanding of intestinal disease, both in neonates and adults, and could lay the groundwork for therapeutic targets.

肠道发育的内吞调节肠粘膜是高度分化的上皮屏障，是维持肠道正常功能的基础。肠。囊泡运输现在被认为是上皮极性、完整性和细胞的重要调节剂。分化，模型系统的研究表明，内吞作用和膜运输是对于肠道的正常个体发生至关重要。在发育过程中，肠上皮细胞在获得成熟的肠道结构之前经历一个强烈的内吞阶段。我们鉴定出内管蛋白（EDTB）是内吞复合物中高度保守的整合膜蛋白发育中的肠道。在体外，EDTB 调节紧密连接组装和接触介导的抑制的扩散。此外，它还与小 GTPase Rab14 结合，调节 GTPase Rab14 之间的运输。内体和顶端质膜。然而 EDTB 在开发过程中的作用是完全未知。我们已经培育出肠上皮细胞特异性 EDTB 敲除小鼠，并且，在初步数据中，表明发育早期的 EDTB 敲除会导致异常的个体发生新生儿肠道，包括顶端内吞复合物的损失、顶端内吞复合物的细胞内积聚质膜蛋白和异常的刷状缘。我们将使用这个 EDTB 条件淘汰赛小鼠以及肠样培养物和细胞系，以确定这些变化的机制基础在上皮分化中。我们将定义与紧密连接蛋白相互作用的机制定义调解其影响的 EDTB 领域。另外，我们将使用 Rab14 来分析 Rab14 的作用肠类肠培养物和人肠上皮细胞系。最后，我们将检查信号已知影响肠道分化并定义其与 EDTB 和顶端相互作用的途径内体。总的来说，这些研究将深入了解肠道的细胞机制。发育，这对于我们了解肠道功能至关重要。实验完成后，该提案中概述的将定义囊泡运输在正常生长、发育和体内平衡中的作用肠上皮。这些基础知识将对我们对肠道的理解产生影响。新生儿和成人的疾病，并可以为治疗目标奠定基础。